SUVA as control parameter for the effective ozonation of organic pollutants in secondary effluent.

Recent research projects have shown a good suitability of the ozonation process to transform trace concentrations of most pharmaceuticals in wastewater treatment plant (WWTP) effluents. The concentrations of carbamazepine and 17 alpha-ethinylestradiol, for instance, were reduced below their detection limits by use of ozone dosages resulting in a specific ozone consumption of 0.5 mg O3/mg DOC0. At the same time a good disinfection performance was achieved. The given hygienic requirements of the EU bathing water directive (e.g. 2,000 N/100 mL faecal coliforms) are fulfilled without the formation of bromate (<10 microg/L). As technical control parameter of the ozonation process usually the residual ozone in the liquid phase or in the off-gas are used. However, at very low specific ozone consumptions, ozone reacts instantaneously with dissolved compounds and cannot be detected. Hence, alternative parameters should be used for effective operation control. The present paper evaluates the relation between UVA decrease and the removal of different compounds (endocrine disrupting compounds, pharmaceuticals, iodinated X-ray contrast media), microbial parameters and bromate formation. The results can be used as a guideline for the control of the oxidation performance at large scale ozonation units.

Outcomes of a 2-year investigation on enhanced biological nutrients removal and trace organics elimination in membrane bioreactor (MBR).

Two configurations of membrane bioreactors were identified to achieve enhanced biological phosphorus and nitrogen removal, and assessed over more than two years with two parallel pilot plants of 2m3 each. Both configurations included an anaerobic zone ahead of the biological reactor, and differed by the position of the anoxic zone: standard pre-denitrification, or post-denitrification without dosing of carbon source. Both configurations achieved improved phosphorus removal. The goal of 50 microgP/L in the effluent could be consistently achieved with two types of municipal wastewater, the second site requiring a low dose of ferric salt ferric salt < 3 mgFe/L. The full potential of biological phosphorus removal could be demonstrated during phosphate spiking trials, where up to 1 mg of phosphorus was biologically eliminated for 10 mg BOD5 in the influent. The post-denitrification configuration enabled a very good elimination of nitrogen. Daily nitrate concentration as low as 1 mgN/L could be monitored in the effluent in some periods. The denitrification rates, greater than those expected for endogenous denitrification, could be accounted for by the use of the glycogene pool, internally stored by the denitrifying microorganisms in the anaerobic zone. Pharmaceuticals residues and steroids were regularly monitored on the two parallel MBR pilot plants during the length of the trials, and compared with the performance of the Berlin-Ruhleben WWTP. Although some compounds such as carbamazepine were persistent through all the systems, most of the compounds could be better removed by the MBR plants. The influence of temperature, sludge age and compound concentration could be shown, as well as the significance of biological mechanisms in the removal of trace organic compounds.

Enhanced biological phosphorus removal process implemented in membrane bioreactors to improve phosphorous recovery and recycling.

The enhanced biological phosphorus removal (EBPR) process was adapted to membrane bioreactor (MBR) technology. One bench-scale plant (BSP, 200-250 L) and two pilot plants (PPs, 1,000-3,000 L each) were operated under several configurations, including pre-denitrification and post-denitrification without addition of carbon source, and two solid retention times (SRT) of 15 and 26 d. The trials showed that efficient Bio-P removal can be achieved with MBR systems, in both pre- and post-denitrification configurations. EBPR dynamics could be clearly demonstrated through batch-tests, on-line measurements, profile analyses, P-spiking trials, and mass balances. High P-removal performances were achieved even with high SRT of 26 d, as around 9 mgP/L could be reliably removed. After stabilisation, the sludge exhibited phosphorus contents of around 2.4%TS. When spiked with phosphorus (no P-limitation), P-content could increase up to 6%TS. The sludge is therefore well suited to agricultural reuse with important fertilising values. Theoretical calculations showed that increased sludge age should result in a greater P-content. This could not be clearly demonstrated by the trials. This effect should be all the more significant as the influent is low in suspended solids.

Transmission of Trichosporon asahii oesophagitis by a contaminated endoscope.

Two cases of oesophageal trichosporonosis due to a suspected nosocomial infection are reported. Both the patients were immunocompetent and had undergone an endoscopic examination on the same day. Six strains of Trichosporon were isolated: three strains from the oesophageal biopsy of the first patient, one strain from the endoscopic forceps, one from the air in the endoscopy room, and one from the oesophageal biopsy of the second patient. The nosocomial nature of the infection and the role of the endoscopic forceps in transporting the micro-organism was suspected, but the morphology and physiology of the isolated strains did not confirm such hypothesis. To elucidate the nature of the infection and the genetic similarities of the strains isolated, all strains were typed with RFLPs of the rDNA fragment and with RAPD. The results of RAPD using primer (GTG)5 (GACA)4, M13 core sequence, and the 15-mer oligonucleotide GAGGGTGGXGGXTCT indicated the molecular identity of three strains supporting the hypothesis concerning a transport of the aetiological agent from the first patient to the second and that the carrier was the forceps of the endoscopic device.

Molecular subtyping of clinical and environmental strains of Cryptococcus neoformans variety neoformans serotype A isolated from southern Italy.

Analysis of ribosomal DNA (rDNA) restriction fragment-length polymorphism (RFLP) and random amplification of polymorphic DNA (RAPD) was used to investigate the genetic variability and biogeographic distribution of clinical and environmental strains of Cryptococcus neoformans isolated from a limited area of southern Italy, where the selection of a predominant cryptococcal genotype could be expected. All isolates belonged to the species Cr. neoformans variety neoformans serotype A. RFLP analysis of a specific rDNA fragment allowed the distinction of strains of Cr. neoformans from closely related fungal reference species, but neither intraspecies nor intravarieties polymorphism was detected. On the contrary, RAPD fingerprints produced by priming with four different primers [(GTG)5, (GACA)4, M13 core sequence and the 8-mer oligonucleotide (GCGGACGG)] were able to characterize the isolates up to the individual level, indicating the presence of marked heterogeneity among Cr. neoformans serotype A strains in southern Italy.

Genetic relatedness and diversity of Cryptococcus neoformans strains in the Maltese Islands.

The genetic relatedness of clinical and environmental Cryptococcus neoformans strains in the Maltese Islands was investigated by randomly amplified polymorphic DNA fingerprinting with four primers. The clinical strains isolate over the course of 1 year from AIDS patients showed identical fingerprints. The electrophoretic patterns of the two clinical strains were also the most common patterns among the environmental strains, but the patterns among the environmental strains showed a wide variability and no correlation with the site of isolation.

Immunoelectronmicroscopic characterization of monoclonal antibodies (MAbs) against Cryptococcus neoformans.

Three monoclonal antibodies (MAbs) (BA4, BD1, CD6) reacted with Cryptococcus neoformans capsular glucuronoxylomannan (GXM) polysaccharide showing distinctive patterns against four serotypes as revealed by enzyme immunoassay (EIA), dot EIA, and immunofluorescence. Immunoelectron microscopy (IEM) was used to characterize binding sites for the MAbs on the C. neoformans capsule. All three MAbs bound to the capsule of serotype A strains 9104 and 9759. Differences in the intensity of binding to the two serotype A strains could not be explained by capsule diameter. The MAb BA-4 IgM bound well to 9759 (large capsule) and poorly to 9104 (small capsule), whereas MAb BD-1 (IgG-1) bound well to strain 9104 and poorly to strain 9759. Spurr's embedment inactivated the BA-4-binding epitopes in the C. neoformans 9759 capsule, but did not inactivate the ones that bound to BD-1. The epitopes recognized by BA-4 were different than the BD-1-binding determinants. The MAb CD-6 bound to a cytoplasmic precursor of capsular GXM. CD-6 (IgG) stained the capsule, cell wall, and cytoplasm of both C. neoformans tester strains. Competitive binding experiments were conducted. Single immunogold labelling showed that BD-1 inhibited the binding of BA-4, but not vice versa. The interaction between CD-6 and BA-4 resulted in a reciprocal inhibition. Double-labelling experiments showed reciprocal inhibition between BA-4 and each of the IgG MAbs. These MAbs are directed against capsular polysaccharide or its intracellular precursor. None of the MAbs stained C. neoformans cap 67, an acapsular mutant that does not contain GXM.

Characterization of Cryptococcus neoformans capsular glucuronoxylomannan polysaccharide with monoclonal antibodies.

Mice were immunized with Cryptococcus neoformans serotype A capsular glucuronoxylomannan (GXM) conjugated to bovine serum albumin-adipic dihydrazide. Two splenocyte fusions yielded two monoclonal antibodies (MAbs) that were highly reactive in dot enzyme immunoassay, immunofluorescence, and sandwich enzyme immunoassay. The first MAb, BD-1 [immunoglobulin G1 (kappa) [IgG1(kappa)]], was GXM-A and GXM-D specific, whereas the second MAb, BA-4 (IgM), reacted with GXM-A and GXM-B. A third MAb, CD-6 [IgG1(kappa)], originated from mice immunized with O-deacetylated GXM-C-bovine serum albumin and reacted with GXMs of all four serotypes. Two of the MAbs (CD-6 and BD-1) were further characterized with chemically modified GXMs. Removal of glucuronosyl residues completely inhibited the binding of both MAbs, implicating (1----2)-beta-glucuronic acid as a key component of the epitope. Removal of (1----2)-beta-xylosyl residues decreased reactivity to an intermediate extent. O deacetylation led to a measurable decrease but had the least inhibitory effect of the three GXM derivatives tested. The combining site for these two MAbs appears to be a complex antigenic determinant involving more than one glycosidic residue.